Use of maillard reaction inhibitors for the treatment of amyloidosis-based disease

ABSTRACT

PCT No. PCT/GB95/00843 Sec. 371 Date Jan. 16, 1997 Sec. 102(e) Date Jan. 16, 1997 PCT Filed Apr. 13, 1995 PCT Pub. No. WO95/28151 PCT Pub. Date Oct. 26, 1995Inhibitors of the Maillard reaction are useful in the treatment or prophylaxis of amyloidosis-based disease, particularly Alzheimer&#39;s disease. Particular examples include aminoguanidine and amphotericin B.

This invention relates to preparations of use in the prophylaxis and treatment of amyloidosis-based diseases, including amyloid encephalopathies such as non-senile dementias, in particular Alzheimer's disease.

Alzheimer's disease is a debilitating and eventually fatal disease of mature adults, typically in middle age. The causative agent is not yet understood, but it has similarities to other dementias, in particular scrapie in sheep and Creutzfeldt-Jakob and Gerstmann-Straussler-Scheinker syndromes and bovine spongiform encephalopathy (BSE). These diseases are characterised by the histopathological features of neural amyloidogenesis. Particular examples are the paired helical filaments that form the neurofibrillar tangles in Alzheimer's brains and the fibrils found in scrapie and other spongiform encephalopathies. These lesions are derived by modification of normal cellular proteins: tau in the case of Alzheimer's and the prion protein (PrP) in the case of the spongiform encephalopathies. The nature of the modification remains unknown, but leads to amyloid plaques, neuronal degeneration and vacuolation, together with astroglial proliferation.

Similarly, there is no known treatment for these diseases although it has been reported (You et al, Nature, 356, 598-601 (1992)) that the onset of scrapie is delayed in the present of the polyene antibiotic amphotericin B.

We have discovered that amphotericin B unexpectedly acts as an inhibitor for the Maillard reaction. Furthermore we have shown that glycation of soluble tau from Alzheimer's brains leads to the formation of amyloid filaments and that the Maillard reaction inhibitor aminoguanidine inhibits this aggregation. The Maillard reaction comprises the well known degradation of proteins in the presence of reductive sugars such as glucose to form glycosylated proteins which are implicated in the complications of diabetes such as cataract, nephropathy, retinopathy and atherosclerosis. Various classes of compounds are known to exhibit an inhibiting effect on the Maillard reaction and hence to be of use in the treatment or prophylaxis of the complications of diabetes and in diseases related to old age. Thus, for example, EP-A-0 433 679 describes 4-hydroxy-5,8-dioxoquinoline derivatives of the following formulae

(a) a 4-hydroxy-5,8-dioxoquinoline derivative of the formula (I) or a pharmaceutically acceptable salt thereof: ##STR1## wherein R₁ and R₂ are each hydrogen, methyl, trifluoromethyl, carboxy, methoxycarbonyl or ethoxycarbonyl, and R₃ is hydrogen or hydroxy;

(b) a 4,5,8-trihydroxyquinoline derivative of the formula (II) or a pharmaceutically acceptable salt thereof: ##STR2## wherein R₁, R₂ and R₃ are as defined hereinbefore; (c) a 3-oxophenoxazine derivative of the formula (III) or a pharmaceutically acceptable salt thereof: ##STR3## wherein R₄ and R₅ are each hydrogen or form by incorporation of C₁ and C₂ carbon atoms a condensed 2,1-b!pyridine ring optionally substituted with a hydroxyl group and/or a carboxyl group, and R₆ is hydrogen or hydroxy; or

(d) a 3-oxophenoxazine N-oxide of the formula (IV) or a pharmaceutically acceptable salt thereof: ##STR4## wherein R₄, R₅ and R₆ are as defined hereinbefore.

EP-A-0 430 045 similarly discloses ascorbic acid tocopheryl phosphate diesters having Maillard reaction--inhibiting action, namely compounds of the formula: ##STR5## wherein R₁, R₂ and R₃ independently of one another denote hydrogen or methyl, or a pharmaceutically acceptable salt thereof.

Also EP-A-0 325 936 describes aminoguanidine derivatives of the general formula: ##STR6## (wherein, R^(1b) represents carbocyclic or heterocylic ring substituted or unsubstituted by from 1 to 3 group(s) selected from halogen atom, alkyl or alkoxy group of from 1 to 4 carbon atom(s), nitro group, phenoxy group, amino group, hydroxy group and acylamino group of from 2 to 4 carbon atom(s), X_(b) represents single-bond, alkylene group of from 1 to 4 carbon atom(s) or alkenylene group of from 2 to 4 carbon atoms, or R^(1b) together with X_(b) represents alkyl group of from 1 to 4 carbon atom(s), R^(2b) represents hydrogen atom, alkyl group of from 1 to 4 carbon atom(s) or phenyl group substituted or unsubstituted by from 1 to 3 group(s) selected from halogen atom, alkyl or alkoxy group of from 1 to 14 carbon atom(s), hydroxy and nitro group), or an acid addition salt thereof.

According to the present invention we provide the use of a pharmaceutically acceptable Maillard reaction inhibitor in the preparation of a pharmaceutical composition for the treatment or prophylaxis of amyloidosis-based disease, including amyloid encephalopathies such as non-senile dementias, in particular such disease in humans, especially Alzheimer's disease. The invention also provides a method of treatment or prophylaxis of such diseases using the Maillard reaction inhibitors.

The Maillard reaction inhibitors of use may comprise any of the various types mentioned above and also aminoglycosides such as amphotericin B and its analogues, as well as any other pharmaceutically acceptable product which can be shown to inhibit the Maillard reaction in a simple in vitro test.

It is envisaged that the dosage rates of the active materials would be in the same range as those used in the treatment of other conditions, up to a maximum determined by the toxicity of the compound in question.

The following description illustrates the Maillard-inhibitory effect of aminoguanidine and amphotericin B and the evidence for the implication of the Maillard reaction in Alzheimer's paired helical filaments.

EXAMPLE 1

Soluble tau protein isolated from normal or Alzheimer's brains was incubated for 24 hr at 37° C. at a concentration of 1 mg/ml in 10 mM reducing sugar solutions in 50 mM phosphate buffer pH 6.8, 1 mM PMSF and 0.1 mM GTP, in the presence or absence of the Maillard reaction inhibitor aminoguanidine. The formation of protein aggregates was analysed by electron microscopy by negative staining. Only soluble tau isolated from Alzheimer's formed aggregates on glycation and this aggregation was inhibited by aminoguanidine. The aggregates formed on glycation by glucose resembled the amyloid fibrils seen in Alzheimer's brains and the Maillard reaction inhibitor, aminoguanidine, inhibited the formation of these filamentous aggregates.

    ______________________________________             Inhibitor   Normal tau  Alzheimer's tau     Sugar   (25 mM)     (1 mg/ml)   (1 mg/ml)     ______________________________________     None                No aggregates                                     No aggregates     Glucose             No aggregates                                     Filamentous                                     aggregates     Glucose aminoguanidine                         No aggregates                                     No aggregates     Fructose            No aggregates                                     Few aggregates     Fructose             aminoguanidine                         No aggregates                                     No aggregates     ______________________________________

EXAMPLE 2

Bovine serum albumin was dried from solutions containing reducing sugars and incubated at 55° C. for 3 weeks, in the presence or absence of various concentrations of either the Maillard reaction inhibitor aminoguanindine or the antibiotic amphotericin B. The extent of the Maillard reaction was quantified spectrophotometrically by the formation of brown colour. The results showed that the antibiotic amphotericin B, which had previously been shown to delay the onset of scrapie in an animal disease by an unknown mechanism, unexpectedly acted as a Maillard reaction inhibitor similar to aminoguanindine.

    ______________________________________                           Ratio      Absorbance     Sugar    Potential    Sugar:Inhibitor                                      extent of     (25 mM)  Inhibitor    (molar basis)                                      browning     ______________________________________     Glucose  None                    0.709     Glucose  aminoguanidine                           3:1        0.603     Glucose  aminoguanidine                           2:1        0.205     Glucose  aminoguanidine                           1:1        0.212     Glucose  aminoguanidine                           1:2        0.211     Glucose  aminoguanidine                           1:3        0.154     None     aminoguanidine                           1:3        0.186     Fructose None                    2.5     Fructose aminoguanidine                           1:3        0.173     Glucose  None         1:3        0.999     Glucose  amphotericin B                           3:1        0.548     Glucose  amphotericin B                           2:1        0.349     Glucose  amphotericin B                           1:1        0.258     Glucose  amphotericin B                           1:2        0.235     Glucose  amphotericin B                           1:3        0.206     None     amphotericin B          0.205     Fructose None                    2.347     Fructose amphotericin B                           1:3        0.224     ______________________________________ 

We claim:
 1. A method to inhibit the aggregation of amyloid filaments associated with an amyloidosis-based disease comprising administering to a subject for treatment or prophylaxis of said disease an effective amount of a pharmaceutically acceptable Maillard reaction inhibitor.
 2. A method according to claim 1, in which the Maillard reaction inhibitor comprises aminoguanidine.
 3. The method of claim 1 wherein said Maillard reaction inhibitor is selected from the group consisting of(a) a compound of the formula ##STR7## wherein R₁ and R₂ are each hydrogen, methyl, trifluoromethyl, carboxy, methoxycarbonyl or ethoxycarbonyl, and R₃ is hydrogen or hydroxy; (b) a compound of the formula ##STR8## wherein R₁, R₂ and R₃ are as defined for formula (I); (c) a compound of the formula ##STR9## wherein R₄ and R₅ are each hydrogen or form by incorporation of C₁ and C₂ carbon atoms a condensed 2,1-b! pyridine ring optionally substituted with a hydroxyl group and/or a carboxy group, and R₆ is hydrogen or hydroxy; (d) a compound of the formula ##STR10## wherein R₄, R₅ and R₆ are as defined for formula (III); and (e) a compound of the formula ##STR11## wherein R₁, R₂ and R₃ independently of one another denote hydrogen or methyl; and the pharmaceutically acceptable salts of compounds of formulas I-V.
 4. The method of claim 1 wherein said Maillard reaction inhibitor is a compound of the formula ##STR12## wherein R^(1b) represents carbocyclic or heterocyclic ring substituted or unsubstituted by from 1 to 3 group(s) selected from halogen atom, alkyl or alkoxy group of from 1 to 4 carbon atom(s), nitro group; and the pharmaceutically acceptable salts of compounds of formula VI.
 5. The method of claim 1 wherein the amyloidosis-based disease is a nonsenile dementia.
 6. The method of claim 4 wherein said dementia is Alzheimer's Disease.
 7. A method for the treatment or prophylaxis of amyloidoses resulting from the Maillard reaction, with the exception of spongiform encephalopathies, comprising administration to a patient of an effective amount of amphotericin B. 